<meta charset="utf8"></head><body>研究胃肠道动力学和药物对运动影响的方案

<ol>
	<li>Jiang, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Therapeutic+role+of+wuda+granule+in+gastrointestinal+motility+disorder+through+promoting+gastrointestinal+motility+and+decreasing+inflammatory+level.">Therapeutic role of wuda granule in gastrointestinal motility disorder through promoting gastrointestinal motility and decreasing inflammatory level.</a> Front Pharmacol. 14, 1-15 (2023).</li><li>Talley, N. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=What+causes+functional+gastrointestinal+disorders?+A+proposed+disease+model.">What causes functional gastrointestinal disorders? A proposed disease model.</a> Am J Gastroenterol. 115 (1), 41-48 (2020).</li><li>Frazer, C., Hussey, L., Bemker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastrointestinal+motility+problems+in+critically+ill+patients.">Gastrointestinal motility problems in critically ill patients.</a> Crit Care Nurs Clin North Am. 30 (1), 109-121 (2018).</li><li>Waclawikova, B., Codutti, A., Alim, K., El, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gut+microbiota-motility+interregulation:+insights+from+in+vivo,+ex+vivo+and+in+silico+studies.">Gut microbiota-motility interregulation: insights from in vivo, ex vivo and in silico studies.</a> Gut Microbes. 14 (1), 1997296 (2022).</li><li>Kwak, D. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Short-term+probiotic+therapy+alleviates+small+intestinal+bacterial+overgrowth,+but+does+not+improve+intestinal+permeability+in+chronic+liver+disease.">Short-term probiotic therapy alleviates small intestinal bacterial overgrowth, but does not improve intestinal permeability in chronic liver disease.</a> Eur J Gastroenterol Hepatol. 26 (12), 1353-1359 (2014).</li><li>Tang, A. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endothelial+Tlr4+and+the+microbiome+drive+cerebral+cavernous+malformations.">Endothelial Tlr4 and the microbiome drive cerebral cavernous malformations.</a> Nature. 545 (7654), 305-310 (2017).</li><li>Mayer, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gut+feelings:+the+emerging+biology+of+gut-brain+communication.">Gut feelings: the emerging biology of gut-brain communication.</a> Nature Rev Neurosci. 12 (8), 453-466 (2011).</li><li>Singh, R., Zogg, H., Ghoshal, U. C., Ro, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Current+treatment+options+and+therapeutic+insights+for+gastrointestinal+dysmotility+and+functional+gastrointestinal+disorders.">Current treatment options and therapeutic insights for gastrointestinal dysmotility and functional gastrointestinal disorders.</a> Front Pharmacol. 13, 1-20 (2022).</li><li>Sanger, G. J., Alpers, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+drugs+for+gastrointestinal+motor+disorders:+Translating+science+to+clinical+need.">Development of drugs for gastrointestinal motor disorders: Translating science to clinical need.</a> Neurogastroenterol Motil. 20 (3), 177-184 (2008).</li><li>Valentin, N., Acosta, A., Camilleri, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+investigational+therapeutics+for+gastrointestinal+motility+disorders:+From+animal+studies+to+phase+ii+trials.">Early investigational therapeutics for gastrointestinal motility disorders: From animal studies to phase ii trials.</a> Expert Opin Investig Drugs. 24 (6), 769-779 (2015).</li><li>Shoor, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Athletes,+nonsteroidal+anti-inflammatory+drugs,+coxibs,+and+the+gastrointestinal+tract.">Athletes, nonsteroidal anti-inflammatory drugs, coxibs, and the gastrointestinal tract.</a> Curr Sports Med Rep. 1 (2), 107-115 (2002).</li><li>Lacy, B. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+antidepressants+on+gastric+function+in+patients+with+functional+dyspepsia.">Effects of antidepressants on gastric function in patients with functional dyspepsia.</a> Am J Gastroenterol. 113 (2), 216-224 (2018).</li><li>Lin, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effects+of+ginsenosides+on+contractile+activity+of+antibiotic-treated+isolated+small+intestinal+smooth+muscle+in+mice.">The effects of ginsenosides on contractile activity of antibiotic-treated isolated small intestinal smooth muscle in mice.</a> Lishizhen Medicine and Materia Medica Research. 33 (04), 790-793 (2022).</li><li>Jespersen, B., Tykocki, N. R., Watts, S. W., Cobbett, P. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Measurement+of+smooth+muscle+function+in+the+isolated+tissue+bath-applications+to+pharmacology+research.">Measurement of smooth muscle function in the isolated tissue bath-applications to pharmacology research.</a> J Vis Exp. (95), e52324 (2015).</li><li>Park, K. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ex+vivo+assessment+of+contractility,+fatigability+and+alternans+in+isolated+skeletal+muscles.">Ex vivo assessment of contractility, fatigability and alternans in isolated skeletal muscles.</a> J Vis Exp. (69), e4198 (2012).</li><li>Semenov, I., Herlihy, J. T., Brenner, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+measurements+of+tracheal+constriction+using+mice.">In vitro measurements of tracheal constriction using mice.</a> J Vis Exp. (64), e3703 (2012).</li><li>Han, J., Chen, X. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Study+overview+of+invisible+spectro+intestine+experiment.">Study overview of invisible spectro intestine experiment.</a> Guiding Journal of Traditional Chinese Medicine and Pharmacy. 14 (3), 94-96 (2008).</li><li>Hao, F. F., Liu, W. Q., Wang, J. N., Nie, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+study+on+the+effect+of+Forsythia+suspensa+water+extract+on+the+movement+of+isolated+intestinal+tract+in+rats.">Experimental study on the effect of Forsythia suspensa water extract on the movement of isolated intestinal tract in rats.</a> Shandong Journal of Traditional Chinese Medicine. 37 (1), 63-66 (2018).</li><li>Ruoff, H. J., Fladung, B., Demol, P., Weihrauch, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastrointestinal+receptors+and+drugs+in+motility+disorders.">Gastrointestinal receptors and drugs in motility disorders.</a> Digestion. 48 (1), 1-17 (1991).</li><li>Foong, D., Zhou, J., Zarrouk, A., Ho, V., O'connor, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Understanding+the+biology+of+human+interstitial+cells+of+cajal+in+gastrointestinal+motility.">Understanding the biology of human interstitial cells of cajal in gastrointestinal motility.</a> Int J Mol Sci. 21 (12), 4540 (2020).</li><li>M&#246;ssner, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Motilit&#228;tsst&#246;rungen+des+gastrointestinaltrakts.">Motilit&#228;tsst&#246;rungen des gastrointestinaltrakts.</a> Der Internist. 56 (6), 613-614 (2015).</li><li>Yin, J., Chen, J. D. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastrointestinal+motility+disorders+and+acupuncture.">Gastrointestinal motility disorders and acupuncture.</a> Autonomic Neuroscience. 157 (1-2), 31-37 (2010).</li></ol>

<table><tbody><tr><td>Acetylcholine&amp;nbsp;</td><td>Sigma, USA</td><td>A6625</td><td></td></tr><tr><td>atropine</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>IA06501</td><td></td></tr><tr><td>Barium chloride</td><td>Macklin Biochemical Co.,Ltd.,Shanghai, China</td><td>B861682</td><td></td></tr><tr><td>CaCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A501330</td><td></td></tr><tr><td>D-glucose</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A610219</td><td></td></tr><tr><td>drawing software</td><td>GraphPad Software, San Diego, California, USA</td><td>&amp;mdash;</td><td></td></tr><tr><td>Epinephrine</td><td>Sigma, USA</td><td>E4642</td><td></td></tr><tr><td>HEPES</td><td>Xiya Reagent Co., Ltd., Shandong, China</td><td>S3872</td><td></td></tr><tr><td>In vitro tissue perfusion system</td><td>PowerLab, ADInstruments, Australia</td><td>ML0146</td><td></td></tr><tr><td>KCl</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100395</td><td></td></tr><tr><td>KH&lt;sub&gt;2&lt;/sub&gt;PO&lt;sub&gt;4&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100781</td><td></td></tr><tr><td>LabChart Professional version 8.3&amp;nbsp;</td><td>ADInstruments, Australia</td><td>&amp;mdash;</td><td></td></tr><tr><td>MgCl&lt;sub&gt;2&lt;/sub&gt;&amp;middot;6H&lt;sub&gt;2&lt;/sub&gt;O</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100288</td><td></td></tr><tr><td>NaCl</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100241</td><td></td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;</td><td>Sangon Biotech Co., Ltd., Shanghai, China</td><td>A100865</td><td></td></tr><tr><td>nifedipine</td><td>Macklin Biochemical Co.,Ltd.,Shanghai, China</td><td>N5087</td><td></td></tr><tr><td>statistical analysis software</td><td>GraphPad Software, San Diego, California, USA</td><td>&amp;mdash;</td><td></td></tr><tr><td>Surgical sutures</td><td>Johnson, USA</td><td>&amp;mdash;</td><td></td></tr></tbody></table>

using an in vitro tissue perfusion system to detect the functional activities of isolated intestinal tubes in real time

<meta charset="utf8"></head><body>作者聚焦:药物对大鼠肠管影响的可靠且可重复的体外评估

使用体外组织灌注系统实时检测离体肠管的功能活动

Gastrointestinal diseases, which have a high incidence, pose considerable challenges for humans. The small intestine is integral to food and drug ...

using-an-vitro-tissue-perfusion-system-to-detect-functional

Research

JoVE Journal

Medicine

Using An In Vitro Tissue Perfusion System to Detect the Functional Activities of Isolated Intestinal Tubes in Real Time

395 次观看.  Chengdu University of Traditional Chinese Medicine. 我们提出了一种分离大鼠肠管并评估药物对体外张力、频率和振幅的影响的方法。这种方法为研究肠管的研究人员提供了一种有价值的方法。当前的实验挑战是保持技术方法的一致性和稳定性，以确保可靠和可重复的科学研究结果，特别是考虑到生物系统固有的敏感性和可变性。该方案具有简单、经济可行、易于控制实验条件、影响因素最小、...

视频: 作者聚焦:药物对大鼠肠管影响的可靠且可重复的体外评估

Real-time Measurement of Epithelial Barrier Permeability in Human Intestinal Organoids

Advances in 3D culture of intestinal tissues obtained through biopsy or generated from pluripotent stem cells via directed differentiation, have resulted in sophisticated in vitro models of the intestinal mucosa. Leveraging these emerging model systems will require adaptation of tools and techniques developed for 2D culture systems and animals. Here, we describe a technique for measuring epithelial barrier permeability in human intestinal organoids in real-time. This is accomplished by microinjection of fluorescently-labeled dextran and imaging on an inverted microscope fitted with epifluorescent filters. Real-time measurement of the barrier permeability in intestinal organoids facilitates the generation of high-resolution temporal data in human intestinal epithelial tissue, although this technique can also be applied to fixed timepoint imaging approaches. This protocol is readily adaptable for the measurement of epithelial barrier permeability following exposure to pharmacologic agents, bacterial products or toxins, or live microorganisms. With minor modifications, this protocol can also serve as a general primer on microinjection of intestinal organoids and users may choose to supplement this protocol with additional or alternative downstream applications following microinjection.

This protocol describes the measurement of epithelial barrier permeability in real-time following pharmacologic treatment in human intestinal organoids using fluorescent microscopy and live cell microscopy.

人肠道 Organoids 上皮屏障通透性的实时测量

Advances in 3D culture of intestinal tissues obtained through biopsy or generated from pluripotent stem cells via directed differentiation, have resulted ...

科研

发育生物学

Perfusable Vascular Network with a Tissue Model in a Microfluidic Device

A spheroid (a multicellular aggregate) is regarded as a good model of living tissues in the human body. Despite the significant advancement in the spheroid cultures, a perfusable vascular network in the spheroids remains a critical challenge for long-term culture required to maintain and develop their functions, such as protein expressions and morphogenesis. The protocol presents a novel method to integrate a perfusable vascular network within the spheroid in a microfluidic device. To induce a perfusable vascular network in the spheroid, angiogenic sprouts connected to microchannels were guided to the spheroid by utilizing angiogenic factors from human lung fibroblasts cultured in the spheroid. The angiogenic sprouts reached the spheroid, merged with the endothelial cells co-cultured in the spheroid, and formed a continuous vascular network. The vascular network could perfuse the interior of the spheroid without any leakage. The constructed vascular network may be further used as a route for supply of nutrients and removal of waste products, mimicking blood circulation in vivo. The method provides a new platform in spheroid culture toward better recapitulation of living tissues.

The protocol describes how to engineer a perfusable vascular network in a spheroid. The spheroid's surrounding microenvironment is devised to induce angiogenesis and connect the spheroid to the microchannels in a microfluidic device. The method allows the perfusion of the spheroid, which is a long-awaited technique in three-dimensional cultures.

微流控装置中组织模型的 Perfusable 血管网

A spheroid (a multicellular aggregate) is regarded as a good model of living tissues in the human body. Despite the significant advancement in the ...

生物工程

Ex Vivo Intestinal Sacs to Assess Mucosal Permeability in Models of Gastrointestinal Disease

The epithelial barrier is the first innate defense of the gastrointestinal tract and selectively regulates transport from the lumen to the underlying tissue compartments, restricting the transport of smaller molecules across the epithelium and almost completely prohibiting epithelial macromolecular transport. This selectivity is determined by the mucous gel layer, which limits the transport of lipophilic molecules and both the apical receptors and tight junctional protein complexes of the epithelium. In vitro cell culture models of the epithelium are convenient, but as a model, they lack the complexity of interactions between the microbiota, mucous-gel, epithelium and immune system. On the other hand, in vivo assessment of intestinal absorption or permeability may be performed, but these assays measure overall gastrointestinal absorption, with no indication of site specificity. Ex vivo permeability assays using &#34;intestinal sacs&#34; are a rapid and sensitive method of measuring either overall intestinal integrity or comparative transport of a specific molecule, with the added advantage of intestinal site specificity. Here we describe the preparation of intestinal sacs for permeability studies and the calculation of the apparent permeability (Papp) of a molecule across the intestinal barrier. This technique may be used as a method of assessing drug absorption, or to examine regional epithelial barrier dysfunction in animal models of gastrointestinal disease.

Ex Vivo Intestinal Sacs to Assess Mucosal Permeability in Models of Gastrointestinal Disease

This protocol describes the use of excised intestinal tissue preparations or "intestinal sacs" as an ex vivo model of intestinal barrier function. This model may be used to assess integrity of both the epithelial barrier and the mucous gel layer at specific intestinal sites in animal models of digestive disease.

体外肠囊评估黏膜通透性在胃肠道疾病模型

The epithelial barrier is the first innate defense of the gastrointestinal tract and selectively regulates transport from the lumen to the underlying ...

医学

用于灌注血管离体培养的新型 3D 打印生物反应器

Ex Vivo Perfusion Culture of Large Blood Vessels in a 3D Printed Bioreactor

Vascular disease forms the basis of most cardiovascular diseases (CVDs), which remain the primary cause of mortality and morbidity worldwide. Efficacious surgical and pharmacological interventions to prevent and treat vascular disease are urgently needed. In part, the shortage of translational models limits the understanding of the cellular and molecular processes involved in vascular disease. Ex vivo perfusion culture bioreactors provide an ideal platform for the study of large animal vessels (including humans) in a controlled dynamic environment, combining the ease of in vitro culture and the complexity of the live tissue. Most bioreactors are, however, custom manufactured and therefore difficult to adopt, limiting the reproducibility of the results. This paper presents a 3D printed system that can be easily produced and applied in any biological lab, and provides a detailed protocol for its setup, enabling users&#39; operation. This innovative and reproducible ex vivo perfusion culture system enables the culture of blood vessels for up to 7 days in physiological conditions. We expect that adopting a standardized perfusion bioreactor will support a better understanding of physiological and pathological processes in large animal blood vessels and accelerate the discovery of new therapeutics.

作者聚焦:EasyFlow - 一种适用于大型血管培养的经济且适应性强的灌流生物反应器 

This protocol presents the setup and operation of a newly developed, 3D printed bioreactor for the ex vivo culture of blood vessels in perfusion. The system is designed to be easily adopted by other users, practical, affordable, and adaptable to different experimental applications, such as basic biology and pharmacological studies.

离体 在3D打印生物反应器中对大血管进行灌注培养

Vascular disease forms the basis of most cardiovascular diseases (CVDs), which remain the primary cause of mortality and morbidity worldwide. Efficacious ...

Canine Intestinal Organoids in a Dual-Chamber Permeable Support System

The permeable support system is typically used in conjunction with traditional two-dimensional (2D) cell lines as an in vitro tool for evaluating the oral permeability of new therapeutic drug candidates. However, the use of these conventional cell lines has limitations, such as altered expression of tight junctions, partial cell differentiation, and the absence of key nuclear receptors. Despite these shortcomings, the Caco-2 and MDCK models are widely accepted and validated for the prediction of human in vivo oral permeability.
Dogs are a relevant translational model for biomedical research due to their similarities in gastrointestinal anatomy and intestinal microflora with humans. Accordingly, and in support of parallel drug development, the elaboration of an efficient and accurate in vitro tool for predicting in vivo drug permeability characteristics both in dogs and humans is highly desirable. Such a tool could be the canine intestinal organoid system, characterized by three-dimensional (3D), self-assembled epithelial structures derived from adult stem cells.
The (1) Permeable Support Seeding Protocol describes the experimental methods for dissociating and seeding canine organoids in the inserts. Canine organoid isolation, culture, and harvest have been previously described in a separate set of protocols in this special issue. Methods for general upkeep of the canine intestinal organoid monolayer are discussed thoroughly in the (2) Monolayer Maintenance Protocol. Additionally, this protocol describes methods to assess the structural integrity of the monolayer via transepithelial electrical resistance (TEER) measurements and light microscopy. Finally, the (3) Permeability Experimental Protocol&#160;describes the tasks directly preceding an experiment, including in vitro validation of experimental results.
Overall, the canine organoid model, combined with a dual-chamber cell culture technology, overcomes limitations associated with 2D experimental models, thereby improving the reliability of predictions of the apparent oral permeability of therapeutic drug candidates both in the canine and human patient.

Here, we present a protocol describing the culture of canine intestinal organoids in a dual-chamber, permeable support system. Organoid seeding in the permeable supports, the monolayer maintenance, and subsequent drug permeability experiments are described.

双室渗透支持系统中的犬肠类器官

The permeable support system is typically used in conjunction with traditional two-dimensional (2D) cell lines as an in vitro tool for evaluating the oral ...

免疫与感染

Investigating Intestinal Barrier Breakdown in Living Organoids

Organoids and three-dimensional (3D) cell cultures allow the investigation of complex biological mechanisms and regulations in vitro, which previously was not possible in classical cell culture monolayers. Moreover, monolayer cell cultures are good in vitro model systems but do not represent the complex cellular differentiation processes and functions that rely on 3D structure. This has so far only been possible in animal experiments, which are laborious, time consuming, and hard to assess by optical techniques. Here we describe an assay to quantitatively determine the barrier integrity over time in living small intestinal mouse organoids. To validate our model, we applied interferon gamma (IFN-&#947;) as a positive control for barrier destruction and organoids derived from IFN-&#947; receptor 2 knock out mice as a negative control. The assay allowed us to determine the impact of IFN-&#947; on the intestinal barrier integrity and the IFN-&#947; induced degradation of the tight junction proteins claudin-2, -7, and -15. This assay could also be used to investigate the impact of chemical compounds, proteins, toxins, bacteria, or patient-derived probes on the intestinal barrier integrity.

Here we describe a technique to quantify the barrier integrity of small intestinal organoids. The fact that the method is based on living organoids enables the sequential investigation of different barrier integrity modulating substances or combinations thereof in a time-resolved manner.

研究活体器官的肠道屏障分解

Organoids and three-dimensional (3D) cell cultures allow the investigation of complex biological mechanisms and regulations in vitro, which previously was ...

Establishing In Situ Closed Circuit Perfusion of Lower Abdominal Organs and Hind Limbs in Mice

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small size of mouse organs, ex vivo perfusion of bone, bladder, skin, prostate, and reproductive organs is challenging or not feasible. Here, we report for the first time an in situ lower body perfusion circuit in mice that includes the above tissues, but bypasses the main clearance organs (kidney, liver, and spleen). The circuit is established by cannulating the abdominal aorta and inferior vena cava above the iliac artery and vein and cauterizing peripheral blood vessels. Perfusion is performed via a peristaltic pump with perfusate flow maintained for up to 2 h. In situ staining with fluorescent lectin and Hoechst solution confirmed that the microvasculature was successfully perfused. This mouse model can be a very useful tool for studying pathological processes as well as mechanisms of drug delivery, migration/metastasis of circulating tumor cells into/from the tumor, and interactions of immune system with perfused organs and tissues.

A protocol is described for in situ perfusion of the mouse lower body, including the bladder, the prostate, sex organs, bone, muscle and foot skin.

在小鼠中建立下腹部器官和后肢的原点闭路灌注

Ex vivo perfusion is an important physiological tool to study the function of isolated organs (e.g. liver, kidneys). At the same time, due to the small ...

Mouse Fetal Whole Intestine Culture System for Ex Vivo Manipulation of Signaling Pathways and Three-dimensional Live Imaging of Villus Development

Most morphogenetic processes in the fetal intestine have been inferred from thin sections of fixed tissues, providing snapshots of changes over developmental stages. Three-dimensional information from thin serial sections can be challenging to interpret because of the difficulty of reconstructing serial sections perfectly and maintaining proper orientation of the tissue over serial sections. Recent findings by Grosse et al., 2011 highlight the importance of three- dimensional information in understanding morphogenesis of the developing villi of the intestine1. Three-dimensional reconstruction of singly labeled intestinal cells demonstrated that the majority of the intestinal epithelial cells contact both the apical and basal surfaces. Furthermore, three-dimensional reconstruction of the actin cytoskeleton at the apical surface of the epithelium demonstrated that the intestinal lumen is continuous and that secondary lumens are an artifact of sectioning. Those two points, along with the demonstration of interkinetic nuclear migration in the intestinal epithelium, defined the developing intestinal epithelium as a pseudostratified epithelium and not stratified as previously thought1. The ability to observe the epithelium three-dimensionally was seminal to demonstrating this point and redefining epithelial morphogenesis in the fetal intestine. With the evolution of multi-photon imaging technology and three-dimensional reconstruction software, the ability to visualize intact, developing organs is rapidly improving. Two-photon excitation allows less damaging penetration deeper into tissues with high resolution. Two-photon imaging and 3D reconstruction of the whole fetal mouse intestines in Walton et al., 2012 helped to define the pattern of villus outgrowth2. Here we describe a whole organ culture system that allows ex vivo development of villi and extensions of that culture system to allow the intestines to be three-dimensionally imaged during their development.

Mouse Fetal Whole Intestine Culture System for Ex Vivo Manipulation of Signaling Pathways and Three-dimensional Live Imaging of Villus Development

Improved imaging technology is allowing three-dimensional imaging of organs during development. Here we describe a whole organ culture system that allows live imaging of the developing villi in the fetal mouse intestine.

使用体外组织灌注系统实时检测离体肠管的功能活动

副本

摘要

探索更多视频

此视频中的章节

人肠道 Organoids 上皮屏障通透性的实时测量

微流控装置中组织模型的 Perfusable 血管网

体外肠囊评估黏膜通透性在胃肠道疾病模型

离体在3D打印生物反应器中对大血管进行灌注培养

离体在3D打印生物反应器中对大血管进行灌注培养

离体在3D打印生物反应器中对大血管进行灌注培养

双室渗透支持系统中的犬肠类器官

研究活体器官的肠道屏障分解

在小鼠中建立下腹部器官和后肢的原点闭路灌注

老鼠胎儿整肠文化系统

使用体外组织灌注系统实时检测离体肠管的功能活动

副本

摘要

探索更多视频

此视频中的章节

人肠道 Organoids 上皮屏障通透性的实时测量

微流控装置中组织模型的 Perfusable 血管网

体外肠囊评估黏膜通透性在胃肠道疾病模型

离体 在3D打印生物反应器中对大血管进行灌注培养

离体 在3D打印生物反应器中对大血管进行灌注培养

离体 在3D打印生物反应器中对大血管进行灌注培养

双室渗透支持系统中的犬肠类器官

研究活体器官的肠道屏障分解

在小鼠中建立下腹部器官和后肢的原点闭路灌注

老鼠胎儿整肠文化系统

离体在3D打印生物反应器中对大血管进行灌注培养

离体在3D打印生物反应器中对大血管进行灌注培养

离体在3D打印生物反应器中对大血管进行灌注培养